Journal
TALANTA
Volume 201, Issue -, Pages 419-425Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2019.04.003
Keywords
Rolling circle amplification; Padlock probe; Colorimetric assay; Biosensor; Bst DNA polymerase; Unbuffered ligation
Categories
Funding
- Natural Sciences and Engineering Research Council of Canada (NSERC)
- Consortium de recherche et d'innovation en bioprocedes industriels au Quebec (CRIBIQ)
- Armand-Frappier foundation
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Detection and identification of DNA by PCR has opened tremendous possibilities and allows detection of minute quantities of DNA highly specifically. However, PCR remains confined to laboratory settings because of the need of thermocyclers and other analytical equipment. This led to development of isothermal amplification techniques, among which Pad Lock Probe (PLP)-based Rolling Circle Amplification (RCA) has several advantages, but typically also requires a laboratory apparatus of some sort to measure DNA amplification. To circumvent this limitation, while still taking advantage of PLP-based RCA, we developed a colorimetric assay that relies on pH change. Using this assay, we can detect DNA in the low picomolar range and obtain results observable with the naked eye in only 20 min without any requirement for a thermocycler or other complex device, making it a particularly portable assay.
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